Objective·To explore the biological functions and underlying mechanisms of anaphase-promoting complex subunit 10(ANAPC10)in the development and progression of liver hepatocellular carcinoma(LIHC,often abbreviated as HCC).Methods·By integrating data from The Cancer Genome Atlas(TCGA)_LIHC,the hepatitis B virus-related subgroup(HBV)of the China Hepatocellular Carcinoma Genome Project(CHCC),and the Gene Expression Omnibus(GEO),the expression pattern of ANAPC10 in HCC was analyzed.Western blotting and quantitative real-time PCR(q-PCR)were used to verify the findings in HCC cell lines.shRNA-mediated knockdown of ANAPC10 was performed in MHCC-97H and SNU-398 cell lines to investigate the effect of ANAPC10 depletion on the in vitro proliferation of HCC cells.An orthotopic liver cancer model with Anapc10 knockout was constructed using the hydrodynamic tail-vein injection technique in mice to further confirm the impact of ANAPC10 deficiency in the liver on the development and progression of HCC.Kyoto Encyclopedia of Genes and Genomes(KEGG)and Gene Ontology(GO)enrichment analyses were performed on the RNA-sequencing data from TCGA_LIHC and CHCC_HBV.Results·ANAPC10 was highly expressed in tumor tissues,and its expression level was closely related to patient survival.Downregulation of ANAPC10 in vitro and in vivo effectively inhibited HCC progression.ANAPC10 mainly reprogrammed the metabolism of tumors by affecting the PI3K-AKT-mTOR pathway.In the tumor tissues of the orthotopic liver cancer mouse model in the Anapc10 knockout group,the phosphorylation levels of Akt and S6k were decreased,and changes in the key downstream lipid metabolism proteins Fasn and Scd1 were verified.Conclusion·ANAPC10 is highly expressed in HCC and is positively correlated with poor prognosis.It promotes HCC occurrence and progression by activating the PI3K-AKT-mTOR signaling pathway and enhancing lipid metabolism reprogramming,thereby promoting tumor cell proliferation.These findings expand the understanding of ANAPC10 in tumor progression and suggest potential therapeutic targets for HCC.

Objective:To investigate the expressions of serum cytokines in patients with autoimmune retinopathy (AIR), and their association with disease diagnosis as well as clinical features.Methods:A prospective case-control study was conducted.A total of 90 eyes of 45 AIR patients were consecutively collected in Beijing Tongren Hospital from September 2018 to December 2023.Additionally, age-matched 43 controls (86 eyes) were enrolled, consisting of 21 patients (42 eyes) with retinitis pigmentosa as a disease control group and 22 healthy subjects (44 eyes) as a normal control group.The main clinical outcome measures included best-corrected visual acuity (BCVA), mean deviation (MD) of visual field, central retinal thickness and the maximal response amplitude and implicit time of full-field electroretinography (ff-ERG). A total of 21 serum cytokines were detected by Luminex multiple cytokines assay or ELISA.Differences in serum cytokine concentrations of among groups, the correlation between cytokine levels and clinical outcomes, and the contribution of cytokines to the diagnosis of AIR were analyzed.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Beijing Tongren Hospital (No.TRECKY2018-048). Written informed consent was obtained from each subject.Results:In AIR patients, the expression levels of Th1-type cytokines or receptors (including interferon-γ[IFN-γ], interleukin [IL]-8, C-X-C motif chemokine ligand [CXCL]9, and CXCL10) and Th17-type cytokines (including IL-17 and IL-6) were elevated.There were statistically significant overall differences in pro-inflammatory cytokines/chemokines (IL-6, IL-8, CXCL10, IL-10, IFN-γ, IL-17, C-X3-C motif chemokine ligand 1 [CX3CL]1, CXCL9) among the three groups ( H=10.823, 10.816, 9.633, 10.103, 23.670, 16.493, 9.050, 9.253; all P<0.05). Compared with the disease control group and the healthy control group, the IFN-γ level was significantly increased in the AIR group (both P=0.001). Compared with the healthy control group, the serum levels of IL-6, IL-8, CXCL10, IL-10, IL-17, and CXCL9 in the AIR group were significantly elevated (all P<0.05). The serum level of CX3CL1 was significantly higher in the AIR group than in the disease control group ( P=0.039). The serum level of IFN-γ was significantly higher in the AIR group than in the healthy control and disease control groups, but no association with AIR diagnosis was found ( OR=1.402, 95% CI: 0.710-2.870, P=0.245). Multilevel mixed-effects regression modeling analysis showed that the serum levels of IL-8 and CXCL9 in AIR patients were positively correlated with LogMAR BCVA ( β=0.028, P=0.033; β=0.023, P=0.003). The C-C motif chemokine ligand 11 level was positively correlated with implicit time of ERG a-wave ( β=13.950, P<0.001). The tumor necrosis factor-α level was positively correlated with MD value of visual field ( β=6.310, P=0.002). Granulocyte-macrophage colony-stimulating factor was negatively correlated with the amplitude of ERG a-wave and granulocyte colony-stimulating factor was negatively correlated with ERG b-wave amplitude ( β=-152.700, P<0.001; β=-14.790, P=0.003). Conclusions:Serum Th1 and Th17-type cytokines/chemokines may be involved in the pathogenesis of AIR and are related to disease severity.

Objective:To develop and validate a prediction model for medication adherence among patients receiving allergen sublingual immunotherapy (SLIT).Methods:A prospective cross-sectional study was conducted, and a total of 288 patients who received SLIT treatment at an allergy center in the First Affiliated Hospital with Nanjing Medical University (Jiangsu Province Hospital) from December 2023 to July 2024 were assigned to the modeling group. Additionally, 122 patients from August to October 2024 were assigned to the validation group. Data of patients′ general information, medication beliefs, anxiety levels, social support, disease perception, and medication adherence were collected. Single-factor analysis and LASSO regression were utilized to identify potential predictors, and a prediction model for medication adherence was constructed using multifactorial logistic regression. A nomogram was then developed based on the model. The model′s discriminatory ability was evaluated using receiver operating characteristic curve (ROC), the area under curve (AUC), sensitivity, and specificity. The model was then validated in the validation cohort.Results:Single-factor analysis and LASSO regression identified a total of nine predictive factors. Logistic regression revealed that medical belief tendency [ OR (95% CI) =2.420 (1.116-5.248), P=0.025], the somatic control dimension in self-rating anxiety scales [ OR (95% CI)=1.404 (1.241-1.589), P<0.001], the subjective support dimension in social support assessment [ OR (95% CI)=0.784 (0.725-0.847), P<0.001], and the cognitive dimension in illness perception [ OR (95% CI)=0.725 (0.647-0.813), P<0.001] were independent predictors of medication adherence in patients undergoing SLIT. The AUC value of the model was 0.899 (95% CI=0.863-0.934) in the modeling group and 0.882 (95% CI=0.820-0.944) in the validation group, indicating good discriminatory ability. The optimal cutoff value of the model was 0.493, with a sensitivity of 81.1% and specificity of 85.7% in the modeling group, and a sensitivity of 87.3% and specificity of 82.5% in the validation group. Conclusion:The medication adherence prediction model developed in this study for patients undergoing SLIT exhibits good predictive performance and provides valuable guidance for early intervention by clinical healthcare professionals.

Objective·To explore the function and potential mechanism of suppressor of zeste 12(SUZ12)in hepatocellular carcinoma(HCC).Methods·The expression of SUZ12 in HCC patients was analyzed using R language in liver cancer datasets,and relevant survival curves were drawn.Stable knockdown of SUZ12 was established in the liver cancer cell lines LM3 and Huh7.The knockdown efficiency of SUZ12 was assessed using quantitative real-time PCR(qPCR)and Western blotting.Cell proliferation ability was assessed using CCK-8 assay and colony formation assay.Using the hydrodynamic tail vein injection(HTVI)method,Suz12 was knocked out in the livers of fully immunocompetent mice to explore its tumorigenic function in vivo.The molecular mechanism of SUZ12 regulating HCC was explored using The Cancer Genome Atlas(TCGA)database.R language was used to analyze the relationship between SUZ12 and the expression of cancer stem cell(CSC)markers as well as key glycolysis-related genes.Findings were validated in liver cancer cell lines and mouse tumor tissues.Results·The expression of SUZ12 in liver cancer tissues was higher than in adjacent non-tumor tissues,and its expression increased with higher tumor stage.HCC patients with high SUZ12 expression had poorer prognoses.In LM3 and Huh7 liver cancer cell lines,stable knockdown of SUZ12 reduced cell proliferation ability.In the de novo MYC/Trp53-/-mouse liver cancer model,tumor nodule number and size,and tumor burden in liver tissue were reduced after endogenous knockout of Suz12.TCGA analysis showed that high SUZ12 expression in HCC was enriched in multiple tumor proliferation-and metabolism-related pathways.The expression of SUZ12 was positively correlated with CSC markers and key genes in glycolysis pathway.The mRNA levels of CSC markers and key genes in glycolysis pathway were decreased in liver cancer cell lines with stable SUZ12 knockdown and Suz12 knockout mouse HCC tissues.Conclusion·The expression of SUZ12 is significantly increased in HCC patients and is associated with poor prognosis.Stable knockdown of SUZ12 weakens the proliferative ability of liver cancer cells.Knockout of Suz12 in mice in vivo can suppress the occurrence and development of HCC.The high expression of SUZ12 maintains the CSC pool,induces metabolic reprogramming,and promotes the occurrence and progression of HCC.SUZ12 can serve as a potential biomarker for poor prognosis and a novel target for potential therapeutic intervention in HCC.

Objective:To investigate the expressions of serum cytokines in patients with autoimmune retinopathy (AIR), and their association with disease diagnosis as well as clinical features.Methods:A prospective case-control study was conducted.A total of 90 eyes of 45 AIR patients were consecutively collected in Beijing Tongren Hospital from September 2018 to December 2023.Additionally, age-matched 43 controls (86 eyes) were enrolled, consisting of 21 patients (42 eyes) with retinitis pigmentosa as a disease control group and 22 healthy subjects (44 eyes) as a normal control group.The main clinical outcome measures included best-corrected visual acuity (BCVA), mean deviation (MD) of visual field, central retinal thickness and the maximal response amplitude and implicit time of full-field electroretinography (ff-ERG). A total of 21 serum cytokines were detected by Luminex multiple cytokines assay or ELISA.Differences in serum cytokine concentrations of among groups, the correlation between cytokine levels and clinical outcomes, and the contribution of cytokines to the diagnosis of AIR were analyzed.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Beijing Tongren Hospital (No.TRECKY2018-048). Written informed consent was obtained from each subject.Results:In AIR patients, the expression levels of Th1-type cytokines or receptors (including interferon-γ[IFN-γ], interleukin [IL]-8, C-X-C motif chemokine ligand [CXCL]9, and CXCL10) and Th17-type cytokines (including IL-17 and IL-6) were elevated.There were statistically significant overall differences in pro-inflammatory cytokines/chemokines (IL-6, IL-8, CXCL10, IL-10, IFN-γ, IL-17, C-X3-C motif chemokine ligand 1 [CX3CL]1, CXCL9) among the three groups ( H=10.823, 10.816, 9.633, 10.103, 23.670, 16.493, 9.050, 9.253; all P<0.05). Compared with the disease control group and the healthy control group, the IFN-γ level was significantly increased in the AIR group (both P=0.001). Compared with the healthy control group, the serum levels of IL-6, IL-8, CXCL10, IL-10, IL-17, and CXCL9 in the AIR group were significantly elevated (all P<0.05). The serum level of CX3CL1 was significantly higher in the AIR group than in the disease control group ( P=0.039). The serum level of IFN-γ was significantly higher in the AIR group than in the healthy control and disease control groups, but no association with AIR diagnosis was found ( OR=1.402, 95% CI: 0.710-2.870, P=0.245). Multilevel mixed-effects regression modeling analysis showed that the serum levels of IL-8 and CXCL9 in AIR patients were positively correlated with LogMAR BCVA ( β=0.028, P=0.033; β=0.023, P=0.003). The C-C motif chemokine ligand 11 level was positively correlated with implicit time of ERG a-wave ( β=13.950, P<0.001). The tumor necrosis factor-α level was positively correlated with MD value of visual field ( β=6.310, P=0.002). Granulocyte-macrophage colony-stimulating factor was negatively correlated with the amplitude of ERG a-wave and granulocyte colony-stimulating factor was negatively correlated with ERG b-wave amplitude ( β=-152.700, P<0.001; β=-14.790, P=0.003). Conclusions:Serum Th1 and Th17-type cytokines/chemokines may be involved in the pathogenesis of AIR and are related to disease severity.

Objective:To develop and validate a prediction model for medication adherence among patients receiving allergen sublingual immunotherapy (SLIT).Methods:A prospective cross-sectional study was conducted, and a total of 288 patients who received SLIT treatment at an allergy center in the First Affiliated Hospital with Nanjing Medical University (Jiangsu Province Hospital) from December 2023 to July 2024 were assigned to the modeling group. Additionally, 122 patients from August to October 2024 were assigned to the validation group. Data of patients′ general information, medication beliefs, anxiety levels, social support, disease perception, and medication adherence were collected. Single-factor analysis and LASSO regression were utilized to identify potential predictors, and a prediction model for medication adherence was constructed using multifactorial logistic regression. A nomogram was then developed based on the model. The model′s discriminatory ability was evaluated using receiver operating characteristic curve (ROC), the area under curve (AUC), sensitivity, and specificity. The model was then validated in the validation cohort.Results:Single-factor analysis and LASSO regression identified a total of nine predictive factors. Logistic regression revealed that medical belief tendency [ OR (95% CI) =2.420 (1.116-5.248), P=0.025], the somatic control dimension in self-rating anxiety scales [ OR (95% CI)=1.404 (1.241-1.589), P<0.001], the subjective support dimension in social support assessment [ OR (95% CI)=0.784 (0.725-0.847), P<0.001], and the cognitive dimension in illness perception [ OR (95% CI)=0.725 (0.647-0.813), P<0.001] were independent predictors of medication adherence in patients undergoing SLIT. The AUC value of the model was 0.899 (95% CI=0.863-0.934) in the modeling group and 0.882 (95% CI=0.820-0.944) in the validation group, indicating good discriminatory ability. The optimal cutoff value of the model was 0.493, with a sensitivity of 81.1% and specificity of 85.7% in the modeling group, and a sensitivity of 87.3% and specificity of 82.5% in the validation group. Conclusion:The medication adherence prediction model developed in this study for patients undergoing SLIT exhibits good predictive performance and provides valuable guidance for early intervention by clinical healthcare professionals.

Objective·To explore the biological functions and underlying mechanisms of anaphase-promoting complex subunit 10(ANAPC10)in the development and progression of liver hepatocellular carcinoma(LIHC,often abbreviated as HCC).Methods·By integrating data from The Cancer Genome Atlas(TCGA)_LIHC,the hepatitis B virus-related subgroup(HBV)of the China Hepatocellular Carcinoma Genome Project(CHCC),and the Gene Expression Omnibus(GEO),the expression pattern of ANAPC10 in HCC was analyzed.Western blotting and quantitative real-time PCR(q-PCR)were used to verify the findings in HCC cell lines.shRNA-mediated knockdown of ANAPC10 was performed in MHCC-97H and SNU-398 cell lines to investigate the effect of ANAPC10 depletion on the in vitro proliferation of HCC cells.An orthotopic liver cancer model with Anapc10 knockout was constructed using the hydrodynamic tail-vein injection technique in mice to further confirm the impact of ANAPC10 deficiency in the liver on the development and progression of HCC.Kyoto Encyclopedia of Genes and Genomes(KEGG)and Gene Ontology(GO)enrichment analyses were performed on the RNA-sequencing data from TCGA_LIHC and CHCC_HBV.Results·ANAPC10 was highly expressed in tumor tissues,and its expression level was closely related to patient survival.Downregulation of ANAPC10 in vitro and in vivo effectively inhibited HCC progression.ANAPC10 mainly reprogrammed the metabolism of tumors by affecting the PI3K-AKT-mTOR pathway.In the tumor tissues of the orthotopic liver cancer mouse model in the Anapc10 knockout group,the phosphorylation levels of Akt and S6k were decreased,and changes in the key downstream lipid metabolism proteins Fasn and Scd1 were verified.Conclusion·ANAPC10 is highly expressed in HCC and is positively correlated with poor prognosis.It promotes HCC occurrence and progression by activating the PI3K-AKT-mTOR signaling pathway and enhancing lipid metabolism reprogramming,thereby promoting tumor cell proliferation.These findings expand the understanding of ANAPC10 in tumor progression and suggest potential therapeutic targets for HCC.

Objective·To explore the function and potential mechanism of suppressor of zeste 12(SUZ12)in hepatocellular carcinoma(HCC).Methods·The expression of SUZ12 in HCC patients was analyzed using R language in liver cancer datasets,and relevant survival curves were drawn.Stable knockdown of SUZ12 was established in the liver cancer cell lines LM3 and Huh7.The knockdown efficiency of SUZ12 was assessed using quantitative real-time PCR(qPCR)and Western blotting.Cell proliferation ability was assessed using CCK-8 assay and colony formation assay.Using the hydrodynamic tail vein injection(HTVI)method,Suz12 was knocked out in the livers of fully immunocompetent mice to explore its tumorigenic function in vivo.The molecular mechanism of SUZ12 regulating HCC was explored using The Cancer Genome Atlas(TCGA)database.R language was used to analyze the relationship between SUZ12 and the expression of cancer stem cell(CSC)markers as well as key glycolysis-related genes.Findings were validated in liver cancer cell lines and mouse tumor tissues.Results·The expression of SUZ12 in liver cancer tissues was higher than in adjacent non-tumor tissues,and its expression increased with higher tumor stage.HCC patients with high SUZ12 expression had poorer prognoses.In LM3 and Huh7 liver cancer cell lines,stable knockdown of SUZ12 reduced cell proliferation ability.In the de novo MYC/Trp53-/-mouse liver cancer model,tumor nodule number and size,and tumor burden in liver tissue were reduced after endogenous knockout of Suz12.TCGA analysis showed that high SUZ12 expression in HCC was enriched in multiple tumor proliferation-and metabolism-related pathways.The expression of SUZ12 was positively correlated with CSC markers and key genes in glycolysis pathway.The mRNA levels of CSC markers and key genes in glycolysis pathway were decreased in liver cancer cell lines with stable SUZ12 knockdown and Suz12 knockout mouse HCC tissues.Conclusion·The expression of SUZ12 is significantly increased in HCC patients and is associated with poor prognosis.Stable knockdown of SUZ12 weakens the proliferative ability of liver cancer cells.Knockout of Suz12 in mice in vivo can suppress the occurrence and development of HCC.The high expression of SUZ12 maintains the CSC pool,induces metabolic reprogramming,and promotes the occurrence and progression of HCC.SUZ12 can serve as a potential biomarker for poor prognosis and a novel target for potential therapeutic intervention in HCC.

【Objective】 To investigate the effects of total flavone of oldenlandia diffusa (FOD) on the proliferation and apoptosis of hepatocellular carcinoma (HCC) stem cells sorted from Huh7. 【Methods】 Human HCC cell lines Huh7 was cultured in vitro; CD133 positive (CD133⁺) stem cells in Huh7 cell line were sorted by flow cytometry, and stem cell markers such as Nanog, Oct4 and Sox2 were tested by Western blotting. CD133⁺-Huh7 was stimulated by different concentrations (0 μg/mL, 50 μg/mL, 100 μg/mL and 400 μg/mL) of FOD for different time (24 h, 48 h, 72 h and 96 h). CCK8 and plate cell cloning assay were used to detect the effect of FOD on CD133⁺-Huh7 proliferation while Annexin V-PE/7-AAD was used to detect the effect of FOD on CD133⁺-Huh7 apoptosis. Western blotting was used to detect the protein expressions of protein 53 (P53), factor associated suicide-Fas-associating protein with a novel death domain (Fas-FADD), B-cell lymphoma-2 (Bcl-2), Cleaved-Caspase3, and Bcl-2 associated X protein (Bax). 【Results】 More than 95% of stem cells were purified for further experiments. Cell proliferation of CD133⁺-Huh7 was significantly inhibited by FOD, with the significant suppression at the concentration of 100 μg/mL for 72 h compared with negative control group (P<0.05). The apoptosis rate was significantly upregulated than that in the negative control group (P<0.05). The protein expression of Bcl2 decreased while Bax and Cleaved-Caspae3 increased via FAS/FADDD and P53 axis. 【Conclusion】 FOD can significantly inhibit the proliferation and promote the apoptosis of CD133⁺-Huh7.

Background: Dietary fiber is strongly recommended as the basic treatment for functional constipation according to global guidelines. However, a complete evaluation standard for the laxative functional food remains to be improved in China. Aims: To evaluate the efficacy and safety of the laxative function of a compound fructose-oligosaccharide fiber granule, so as to provide evidence-based medical basis for the evaluation of laxative functional food. Methods: In a randomized, double-blind, parallel and controlled trial with placebo as control, 120 subjects with functional constipation were enrolled in 2 clinical research centers in Beijing, and randomly divided into experimental group and control group (60 cases in each group). Subjects in experimental group were given a compound fructose-oligosaccharide fiber granule dissolved in 50 mL water orally, 1 bag (9 g) per day for 2 weeks; while those in control group were given a placebo granule with the same appearance, specification and dosage as the experimental group. The bowel movement frequency per week, defecation status and stool consistency were recorded before and after the test, and the safety tests were completed. Results: After 2 weeks of treatment, the bowel movement frequency in experimental group increased by (1.63±1.57) times per week, the stool consistency assessed by Bristol stool form scale and the difficulty in defecation were also improved as compared with the baseline (all P<0.05). Furthermore, improvements in experimental group were superior to those in control group (all P<0.05). No allergic and other adverse events were reported during the test, and there were no significant changes in blood, urine, stool routine and blood biochemical indices before and after the test. Conclusions: The compound fructose-oligosaccharide fiber granule tested in this study is proved to have laxative effect and is safety for functional constipation. The testing program is scientific and of feasibility, and may provide a methodology basis for human oral administration trials of laxative functional food.